The demand for coniferous trees, such as pines and firs, to make wood products continues to increase. One proposed solution to this problem is to identify individual trees that possess desirable characteristics, such as a rapid rate of growth, and produce numerous, genetically identical clones of the superior trees by somatic cloning.
Somatic cloning is the process of creating genetically identical trees from tree tissue other than the male and female gametes. In one approach to somatic cloning, plant tissue is cultured in an initiation medium which includes hormones, such as auxins and/or cytokinins, that initiate formation of embryogenic cells that are capable of developing into somatic embryos. The embryogenic cells are then further cultured in a maintenance medium that promotes multiplication of the embryogenic cells to form pre-cotyledonary embryos (i.e., embryos that do not possess cotyledons). The multiplied embryogenic cells are then cultured in a development medium that promotes development of colyledonary somatic embryos which can, for example, be placed within artificial seeds and sown in the soil where they germinate to yield conifer seedlings. The seedlings can be transplanted to a growth site for subsequent growth and eventual harvesting to yield lumber, or wood-derived products.
There is a continuing need to improve the efficiency of somatic cloning of conifer embryos in order to increase production of cotyledonary somatic embryos that are capable of germinating to yield pine plants. Preferably, the conifer somatic embryos formed in vitro are physically and physiologically similar, or identical, to conifer zygotic embryos formed in vivo in conifer seeds. The present invention provides methods that address this need with respect to conifers of the genus Pinis. 